Dynamics of protein structure alteration at single molecular level

单分子水平蛋白质结构改变的动力学

• 批准号: 17360395
• 负责人: TAKAGI Masahiro
• 金额: $ 10.36万
• 依托单位: Japan Advanced Institute of Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17360395/
• 关键词: Protein; Structure; Amyloid; Membrane; Interaction; タンパク質; フォールディング; 構造変化; 拡張遺伝子工学; ウシトリプシンインヒビター; ミクログロブリン

(1) In order to perform analysis of protein structural alteration at single molecular level using FRET method We attempted fluorescence labeling of bavine pancreatic tripsin inhibitor (BFTI) using a new technique of four base codon method. Fluorescent proteins were obtained but not enough yield for studies at molecular level(2) The effects of negatively charged phosphatidylserine (PS) and neutral phosphatidylcholine (PC) on the structure of wild-type and mutant bovine pancreatic trypsin inhibitor (BPTI) at neutral pH were investigated. The presence of PC did not have any effect on the protein structure whilst PS induced a non-native structure in three mutant BPTI proteins. However, the negatively charged phospholipid did not have any effect on wild-type BPTI. The findings revealed that (i) elimination of some disulphide bonds, results in dramatic change in protein structure, and, (ii) that this biochemical interaction is surface-driven and that electrostatic interactions may play a ver … More y strong role in influencing the fore-stated changes in protein structure.(3) Patients on long term hemodyalysis frequently suffer from dialysis-related amyloidosis (DRA). DRA is a pathology associated with a persistent increase of monomeric β2- microglobulin (β2-m) caused by dialysis treatment of renal failure. The mechanism of formation of amyloid fibrils for this protein, as for all the other amyloidogenic proteins, is not clear, and there was great uncertainty about the molecular bases of amyloid deposition when the amyloidogenicity of β2-m was discovered. In this study, we investigated formation of variant β2-m in serum of a patient who receives treatment of hemodialysis. Variant β2-m is derived from restricted cleavage of β2-M. The possible factor involved in formation of variant β2-m was also estimated. The following conclusions are proposed. Formation of variant β2-m, namely restricted cleavage of β2-m, might be caused by proteolytic cleavage by a serum protease(s). However, it is not clear whether the variant β2-m can trigger formation of amyloid. Protein aggregation formation is enhanced in serum from a patient. Less

(1)为了利用FRET法在单分子水平上进行蛋白质结构变化的分析，我们尝试使用四碱基密码子法的新技术对牛胰胰蛋白酶抑制剂(BFTI)进行荧光标记。获得了荧光蛋白，但产量不足以进行分子水平的研究(2) 研究了中性pH下带负电荷的磷脂酰丝氨酸(PS)和中性磷脂酰胆碱(PC)对野生型和突变型牛胰蛋白酶抑制剂(BPTI)结构的影响。 PC 的存在对蛋白质结构没有任何影响，而 PS 则在三种突变 BPTI 蛋白质中诱导非天然结构。然而，带负电荷的磷脂对野生型BPTI没有任何影响。研究结果表明，(i) 消除一些二硫键会导致蛋白质结构发生巨大变化，(ii) 这种生化相互作用是表面驱动的，静电相互作用可能在影响上述蛋白质结构变化方面发挥非常重要的作用。(3) 长期血液透析患者经常患有透析相关淀粉样变性 (DRA)。 DRA 是一种与肾衰竭透析治疗引起的单体 β2-微球蛋白 (β2-m) 持续增加相关的病理学。与所有其他淀粉样蛋白形成蛋白一样，该蛋白形成淀粉样蛋白原纤维的机制尚不清楚，并且当β2-m的淀粉样蛋白形成性被发现时，淀粉样蛋白沉积的分子基础存在很大的不确定性。在这项研究中，我们研究了接受血液透析治疗的患者血清中变体 β2-m 的形成。变体β2-m源自β2-M的限制性裂解。还估计了参与变体β2-m形成的可能因素。提出以下结论。变体β2-m的形成，即β2-m的限制性切割，可能是由血清蛋白酶的蛋白水解切割引起的。然而，尚不清楚变体β2-m是否可以触发淀粉样蛋白的形成。患者血清中蛋白质聚集的形成增强。较少的

项目成果

Studies about stability of membrane raft domain

膜筏结构域稳定性研究

• 发表时间: 2007
• 作者: Yamaguchi;K.;et. al.
• 通讯作者: et. al.

光による脂質二分子膜小胞の形態転移

光诱导脂质双层囊泡的形状转变

• 发表时间: 2007
• 作者: 石井健一;長崎健;浜田勉;高木昌宏
• 通讯作者: 高木昌宏

Endoplasmic reticulum stress responses as criteria to evaluate functional foods and cosmetics

内质网应激反应作为评价功能性食品和化妆品的标准

• 发表时间: 2007
• 作者: M. Takagi;K. Yamaguchi;T. Hamada (Invited)
• 通讯作者: T. Hamada (Invited)

生体モデル膜ドメインの出芽ダイナミクス

生物模型膜域的萌芽动力学

• 发表时间: 2007
• 作者: 浜田勉;高木昌宏;三浦陽子
• 通讯作者: 三浦陽子

Stretched-exponential analysis of heat-induced aggregation of apo-concanavalin A.

脱辅基刀豆球蛋白 A 热诱导聚集的拉伸指数分析。

• 发表时间: 2005
• 期刊: Protein J. 24
• 作者: J.Ishii;A.Kondo et al.;Y.Ito;S.Shbasaki et al.;T.Fukusaki et al.;T.Konishi et al.;T.Fukuda et al.;J.K.Kim et al.;M.Kudo et al.
• 通讯作者: M.Kudo et al.